Amit Pratush » Публикация

Поделиться публикацией:
Опубликовать в блог:
Опубликовано 2013-10-14 Опубликовано на SciPeople2013-10-14 14:44:30 ЖурналAppl Biochem Biotechnol

Cloning, sequencing and expression of NHase gene of mutant strain of Rhodococcus rhodochrous PA-34 in E.coli
Pratush A, Seth A and Bhalla TC / Amit Pratush
Аннотация The NHase encoding gene of mutant 4D was isolated by PCR amplification. The NHase gene of mutant 4D was successfully cloned and expressed in Escherichia coli by using Ek/LIC Duet cloning kits (Novagen). For the active expression of the NHase gene, the co-expression of small cobalt transporter gene (P-protein gene) has also been co-expressed with NHase gene E. coli. The nucleotide sequence of this NHase gene revealed high homology with the H-NHase of Rhodococcus rhodochrous J1. The recombinant E. coli cells showed higher NHase activity (5.9 U/mg dcw) as compared to the wild (4.1 U/mg dcw) whereas it is less than the mutant strain (8.4 U/mg dcw). Addition of cobalt ion in Luria– Bertani medium is needed up to a very small concentration (0.4 mM) for NHase activity. The recombinant E. coli exhibited maximum NHase activity at 6 h of incubation and was purified with a yield of 56 % with specific activity of 37.1 U/mg protein.
Ключевые слова публикации:

Нет комментариев

Вам необходимо зайти или зарегистрироваться для комментирования